重组Anti-Transcription factor AP-2-alpha抗体[EPR2688(2)]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling Transcription factor AP-2-alpha with Purified ab108311 at 1/100 dilution (1.07 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

Intracellular Flow Cytometry analysis of JAR (Human placenta choriocarcinoma epithelial cell) cells labeling Transcription factor AP-2-alpha with Purified ab108311 at 1/20 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

Immunocytochemistry analysis of JAR (Human placenta choriocarcinoma epithelial cell) cells labeling Transcription factor AP-2-alpha with Purified ab108311 at 1/50 dilution (3.4 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IP

Lab

Immunoprecipitation - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

Purified ab108311 at 1/20 dilution (0.5µg) immunoprecipitating Transcription factor AP-2-alpha in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+) : ab108311 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab108311 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/10,000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 48 kDa

All lanes:

Immunoprecipitation - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab108311)

Predicted band size: 48 kDa

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• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse breast tissue sections labeling Transcription factor AP-2-alpha with Purified ab108311 at 1/100 dilution (1.07 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat breast tissue sections labeling Transcription factor AP-2-alpha with Purified ab108311 at 1/100 dilution (1.07 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Lab

Western blot - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

Lanes 1- 2 : Merged signal (red and green). Green - ab108311 observed at 48 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab108311 was shown to react with Transcription factor AP-2-alpha in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265122 (knockout cell lysate ab257736) was used. Wild-type HeLa and TFAP2A knockout HeLa cell lysates were subjected to SDS-PAGE. ab108311 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab108311) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TFAP2A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TFAP2A (Transcription factor AP-2-alpha) knockout HeLa cell line (<a href='/products/cell-lines/human-tfap2a-transcription-factor-ap-2-alpha-knockout-hela-cell-line-ab265122'>ab265122</a>)

Predicted band size: 122 kDa,125 kDa,32 kDa,39 kDa,40 kDa,41 kDa,42 kDa,45 kDa,48 kDa,74 kDa,77 kDa,83 kDa,97 kDa

Observed band size: 100 kDa,120 kDa,125 kDa,150 kDa,31 kDa,40 kDa,41 kDa,42 kDa,45 kDa,48 kDa,50 kDa,70 kDa,74 kDa,75 kDa,77 kDa,95 kDa

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• WB

Lab

Western blot - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

Lanes 1 - 2 : Merged signal (red and green). Green - ab108311 observed at 48 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab108311 was shown to recognize Transcription factor AP-2-alpha in wild-type HAP1 cells as signal was lost at the expected MW in TFAP2A (Transcription factor AP-2-alpha) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and TFAP2A (Transcription factor AP-2-alpha) knockout samples were subjected to SDS-PAGE. ab108311 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab108311) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

TFAP2A (Transcription factor AP-2-alpha) knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 44 kDa,48 kDa

Observed band size: 44 kDa

false

• WB

Unknown

Western blot - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

All lanes:

Western blot - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (ab108311) at 1/10000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg

Lane 3:

Mouse skin lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 48 kDa

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• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 SK-MEL-28 (human malignant melanoma) cells and 5 µg of ab108311 [EPR2688(2)]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 SK-MEL-28 (human malignant melanoma) cells and 5 µg of ab108311 [EPR2688(2)]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Transcription factor AP-2-alpha antibody [EPR2688(2)] (AB108311)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 SK-MEL-28 (human malignant melanoma) cells and 5 µg of ab108311 [EPR2688(2)]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.